The present invention relates to a process for producing a phytosterol (or plant sterol) from a crude fatty acid ester derived from a vegetable (or plant) fat and/or oil.
Phytosterols are not only used in clinical administration etc. expected to have an effect of reducing cholesterols in blood but also utilized generally as a raw or starting material for pharmaceutical agents as substitutes for cholesterols or as emulsifiers or emulsion-stabilizers in cosmetics and foods.
As the process for producing phytosterols, a method in which sterols contained in vegetable oils such as a soybean oil, a rapeseed oil, a rice bran oil, a palm oil, a palm kernel oil and a coconut oil are extracted and purified from a deodorized concentrate etc. is practically used. There is known the extracting method which comprises adding a lower alcohol to a deodorized distillate of fats and oils to esterify with an acid catalyst, then washing with water to remove the catalyst, transesterifying by adding an additional lower alcohol and alkali catalyst, calmly laying overnight the resultant solution after the reaction to precipitate crystals, separating the crystals by the filtration, washing with a small amount of hexane to further re-crystallize (JP-B 52-8309).
In this method, however, it is necessary to repeatedly re-crystallize in order to obtain highly pure sterols. Then, it is difficult to more easily obtain highly pure phytosterols.
The object of the present invention is to provide a process for producing a highly pure phytosterol, at a high yield in particular, by a simple and easy treatment from a crude fatty acid ester derived from a vegetable fat and/or oil.
The invention a process for producing phytosterol, which comprises (A) bringing a crude fatty acid product derived from a vegetable fat and/or oil including phytosterol into contact with a mixed solvent (or solvent mixture) of an organic solvent and water to crystallize the phytosterol and separating the crystals from the mixed solvent; or (B) mixing a crude fatty acid ester derived from a vegetable fat and/or oil including the phytosterol and a fatty acid ester with a lower alcohol, allowing the mixture to stand at a temperature of 1 to 40xc2x0 C. to precipitate crystals including the fatty acid ester and separating the crystals to take the lower alcohol solution including the phytosterol.
In the method (A), part or all of the treated phytosterol is crystallized. The invention process can be obtained by either the method (step) (A) or (B). Alternatively (A) and (B) may be used in combination with each other.
The present invention provides preferably the above-mentioned process which comprises Steps of (A) and which further comprises allowing a mixture of the crude fatty acid ester with a lower alcohol to stand at a temperature of 1 to 40xc2x0 C. in order to precipitate crystals including the fatty acid ester.
In the above shown method (A), phytosterol existing in crude fatty acid esters of vegetable fats and/or oils can be concentrated by crystallizing the phytosterol in a solvent mixture of an organic solvent and water. A concentrated phytosterol-including product can be obtained this way.
In the method (B), phytosterol existing in crude fatty acid esters of vegetable fats and/or oils can be concentrated by crystallizing the fatty acid esters in a lower alcohol at a temperature of 1 to 40xc2x0 C., separating the crystals out and taking the lower alcohol solution including phytosterol. The lower alcohol solution may include remaining fatty acid esters in addition to concentrated phytosterol. A concentrated phytosterol product can be obtained this way.
The invention provides a process for preparing a concentrated phytosterol product, comprising the above shown (A) or (B). (A) and (B) may be continuously repeated to obtained a very concentrated product of phytosterol. (A) and (B) may be used in combination with each other.
The starting material for (A) or (B) may be a natural source of fats and/or oils including phytosterol. It includes phytosterol, fatty acid esters of phytosterol and other fatty acid esters. Alternatively the natural source may be processed in advance to (A) or (B), for example, by treating fatty acid esters by saponification. The saponification may be effected with an alkali and a mixed solvent of a lower alcohol and water. The product of the invention may be saponified this way to obtain a more concentrated phytosterol product.
The phytosterols in the present invention refer to sterols, contained in vegetable fats and oils, such as sitosterol, stigmasterol, campesterol and brassicasterol.
The vegetable fats and oils used here may be one member of a soybean oil, a rapeseed oil, a rice bran oil, a palm oil, a palm kernel oil and a coconut oil or a mixture of two or more members selected therefrom. Among them, a palm kernel oil, a coconut oil or a palm oil is preferable.
As the crude fatty acid ester derived from vegetable fats and oils in the present invention, there may be preferably used a mixture of various compounds including mainly esters of higher fatty acids with lower alcohols such as esters of C6-28 fatty acids with lower alcohols in particular. The composition thereof cannot be specified because the composition is varied depending on the vegetable fats and oils used as the raw material but the composition may also contain monoglycerides, diglycerides, triglycerides, glycerol etc. That is, the crude fatty acid ester is not particularly limited so far as it contains fatty acid esters and phytosterols. For example, there can also be used a crude fatty acid ester obtained by esterifying the above-mentioned deodorized distillate of a vegetable fat and/or oil and then, if necessary, transesterifying with a lower alcohol such as methanol or a raw material obtained by further distilling the said fatty acid ester product according to the technique described in JP-A 8-500598 and increasing the concentration of the phytosterols by distilling a part of the fatty acid ester product formed by the transesterification reaction.
Alternatively, the above-described vegetable fat and/or oil is esterified and then transesterified with a lower alcohol such as methanol, and the fatty ester product thus obtained is distilled to remove distillates, whereby the concentration of the phytosterols can be increased. The raw material concentrated in this manner can also be used. This concentrate contains not only the phytosterols and fatty ester acids but also unreacted monoglycerides, diglycerides and triglycerides having not reacted in the previous transesterification reaction. There can also be used a crude fatty acid ester obtained by further transesterifying the said concentrate.
In general, these raw materials often contain not less than 20% by weight of the esters of the C6-28 fatty acids with the lower alcohols. In the present invention, fatty acid esters at such concentrations are also preferably used.
As the fatty acid in the present invention, there can be utilized any one selected from a member and a mixture of two or more members of linear or branched saturated or unsaturated ones. The lower alcohols include C1-4 alcohols such as methanol, ethanol, isopropanol, n-propanol and a linear or branched butanol. As the lower alcohol, there can be used a member or a mixture of two or more members thereof. Among them, methanol, ethanol or a mixture thereof is preferable and a methanol-containing member is particularly preferable.
Although the content of the phytosterols in the crude fatty acid ester used in the present invention is not particularly limited, the content is preferably not less than 1% by weight (as compared with the mixture thereof excluding lower alcohols such as methanol) and, in order to exhibit an advantageous effect of the present invention, more preferably 3 to 40% by weight.
In Step A in the present invention, the crude fatty acid ester described above are brought into contact with a mixed solvent comprising an organic solvent and water to crystallize phytosterols. As the organic solvent, there is preferably used one or more solvents having a relative dielectric constant (xcex5r) of not less than 17 at 25xc2x0 C. For available easiness; a member or a mixture of two or more members selected from methanol, ethanol and acetone is more preferable and a methanol-containing member is particularly preferable.
The relative dielectric constant of the organic solvent in the present invention referred to the value shown in xe2x80x9cChemical Index Basic-compilation (Kagaku Binran Kisohen)xe2x80x9d 4th edition published by Maruzen Co., Ltd. or xe2x80x9cSolvent (Youzai) Handbookxe2x80x9d published by Kodansha.
Although the amount of the organic solvent in the mixed solvent used in Step A is not particularly limited, the amount is preferably from 20 to 300% by weight as compared with the crude fatty acid ester. The amount of water in the mixed solvent comprising the organic solvent and water is preferably larger for improving a recoverable ratio of the phytosterols, but a too large amount of water is not economical because of a longer time for separating the crystals from the solution. Accordingly, the amount of water is preferably not less than 1% by weight and more preferably 5 to 150% by weight as compared with the organic solvent.
The crystallization in Step A is preferably carried out by adding water to the mixture of the crude fatty acid ester and the organic solvent. If the crystallization temperature is high, the amount of the precipitated crystals is insufficient to lower the recoverable ratio of the phytosterols, so that the temperature is preferably from 0 to 40xc2x0 C. and more preferably from 1 to 20xc2x0 C. The precipitated crystals are separated from the system (or composition) by publicly known techniques such as filtration, centrifugation and decantation.
This method of the crystallization and separation may comprise adding water, then separating into phases in order to separate a lower phase comprising water/organic solvent, extracting and removing the lower phase, and then separating crystals by allowing an upper phase to stand at a temperature in the range of 0 to 40xc2x0 C. or may comprise allowing the solution to stand at a temperature in the range of 0 to 40xc2x0 C., then separating into phases, extracting the lower phase, and then separating crystals from the upper phase. In the method of the crystallization and separation, the solution is separated by adding water to separate into phases, a phase of water/organic solvent as a lower phase is removed by extracting it, and an upper phase is allowed to stand (or kept) at a temperature in the range of 0 to 40xc2x0 C., whereby the crystals may be separated. Alternatively, the solution is kept at a temperature in the range of 0 to 40xc2x0 C. and then separated into phases, then a lower phase is extracted, and the crystals may be separated from an upper phase. In any method, the crystals to be precipitated are present in an oil phase, and the solution may be operated to crystallization and separation as it is or may be operated to crystallization and separation after the solution is separated into phases to remove the lower phase. The temperature for separating into phases is not particularly limited, but it is preferably from 0 to 100xc2x0 C. and more preferably from 0 to 64xc2x0 C. By these operations, the recoverable ratio of the phytosterols can be significantly improved.
In the present invention, an already known solvent-purifying method i.e. re-crystallization method is applied if necessary to the crystallized precipitation after Step A is finished. The solvent used in this step is mainly an organic solvent such as methanol, but a mixed solvent of an organic solvent and water may also be used. Such an already known re-crystallization step may also be carried out before Step A. That is, if the already known re-crystallization step is used in combination with Step A, the order thereof is not particularly limited.
In the present invention, Step B can be combined before or after Step A or the combined steps of Step A with the already known re-crystallization step. By the combination with Step B, highly pure phytosterols can be extracted at a high yield.
In Step B, the mixture of the crude fatty acid ester and the lower alcohol is regulated such that the amount of the lower alcohol is preferably not less than 2% by weight and more preferably 20 to 300% by weight as compared with the crude fatty acid ester. In this case, a lower alcohol may be additionally (or further) added; however, if the lower alcohol in the esterification step or in the transesterification step of a former step or Step A still remains in the reaction system in the desired amount as compared with the crude fatty acid ester, further addition of the lower alcohol is not required. In short, the presence of the lower alcohol is required and the means therefor is not inquired. The lower alcohol used here includes a C1-4 alcohol described above and is preferably methanol, ethanol or a mixture thereof and is particularly preferably a methanol-containing member.
Next, the mixture which comprises the crude fatty acid ester and the lower alcohol and which is regulated as described above is allowed to stand at 1 to 40xc2x0 C. to precipitate crystals. The crystals thus precipitated comprise the esters of the C6-28 fatty acids with the lower alcohol as the main component. These crystals contain little phytosterols. That is, the phytosterols are present as solution components.
The crystallization temperature kept in Step B is very important. Esters of the C6-28 fatty acids with the lower alcohol begin to be precipitated as crystals at 50xc2x0 C. or less. When the temperature for precipitation by crystallizing is high, the amount of precipitation is not sufficient. Namely, there becomes small a contribution in order to improve the purity of the phytosterols. Then, there is lowered the purity of the phytosterols after Step A or the re-crystallization step which is subsequently carried out to the solution from which the crystals have been separated. On the other hand, when the temperature for cooling is too low, the phytosterols are intermixed in the precipitated crystals to lower the recoverable ratio of the phytosterols. Accordingly, the crystallization temperature is controlled in the range of 1 to 40xc2x0 C. and preferably 3 to 20xc2x0 C. Further, the time during which the solution is kept at 1 to 40xc2x0 C. is preferably 0.1 to 2 hours.
Step A in the present invention is carried out after the fatty acid esters in the crude fatty acid ester for use are removed to the utmost, whereby more highly pure phytosterols can be obtained. Accordingly, Step A is most preferably carried out to a filtrate after Step B is carried out.
In Step B of the present invention, further, the precipitated crystals are separated by a publicly known technique as above-mentioned in Step A, whereby the purity of phytosterols after a crystallizing and precipitating treatment being carried out to a rest solution can be significantly improved. That is, an already known solvent-purifying method i.e. crystallizing and precipitating method is applied to the solution from which the precipitated crystals were separated. The solvent used in this step is not particularly limited. For example, methanol, methyl ethyl ketone, ethanol or the like can be used and methanol is particularly preferable.
According to the process of the present invention, highly pure phytosterols can be produced easily and efficiently from a crude fatty acid ester. Further, according thereto, crystals of fatty acid esters can be precipitated highly selectively from the crude fatty acid ester to separate and remove the crystals from the solution containing the phytosterols. Thus, the phytosterols having the significantly improved purity can be obtained without adversely affecting the yield after the crystalline precipitation.